CAJ | 학술논문

目的：通过优化条件培养基用于非牙源性上皮细胞的体外扩增，为进一步开展组织工程牙提供上皮种子细胞来源奠定基础。方法：获取出生后1 d C57BL 鼠原代口腔黏膜上皮细胞，接种于含有 Y27632的上皮条件培养基的培养瓶内。细胞长满后，消化传代，采用条件培养基与3T3饲养层细胞共培养进行扩增并连续传代，观察细胞生长情况及连续传代后的生物学特性，免疫组织化学进行干细胞鉴定。结果：与传统上皮细胞培养方式相比，含 Y27632的条件性培养基加3T3饲养层方式培养的上皮细胞生长迅速，细胞呈立方形或多角形，呈铺路石样排列。免疫组织化学染色表明这些干细胞表达 CK14，SOX2和LGR5。连续传代至第7代后，细胞形态、生长曲线和干细胞标志没有明显变化。结论：含有 Y27632的条件培养基可有效用于体外大量扩增非牙源性上皮细胞，并能保持干细胞生长特性，有望用于组织工程牙的种子细胞来源。
목적：통과우화조건배양기용우비아원성상피세포적체외확증，위진일보개전조직공정아제공상피충자세포래원전정기출。방법：획취출생후1 d C57BL 서원대구강점막상피세포，접충우함유 Y27632적상피조건배양기적배양병내。세포장만후，소화전대，채용조건배양기여3T3사양층세포공배양진행확증병련속전대，관찰세포생장정황급련속전대후적생물학특성，면역조직화학진행간세포감정。결과：여전통상피세포배양방식상비，함 Y27632적조건성배양기가3T3사양층방식배양적상피세포생장신속，세포정립방형혹다각형，정포로석양배렬。면역조직화학염색표명저사간세포표체 CK14，SOX2화LGR5。련속전대지제7대후，세포형태、생장곡선화간세포표지몰유명현변화。결론：함유 Y27632적조건배양기가유효용우체외대량확증비아원성상피세포，병능보지간세포생장특성，유망용우조직공정아적충자세포래원。
Objective:To characterize the role of conditioned culture media on expansion of non-odontogenic epithelial cells in vitro for potential seed cells of further tissue engineering tooth.Methods:The oral mucosa epithelial cells were harvested from 1 day postnatal mouse and cultured to confluency in conditioned media supplemented with Y27632,then passaged and plated onto 3T3 feeder cells in conditioned media with Y27632 for continuous passage.Histology and immunohistochemistry were performed for biological characteris-tics analysis of cells.Results:Compared with the traditional ways of epithelial cell culture,the oral mucosa epithelial cells plated onto 3T3 feeder cells in conditioned media supplemented with Y27632 grew rapidly,which showed a cuboidal,polygonal or pavement-like shape and expressed CK14,SOX2 and LGR5.After continuous passage,the cell morphology,growth curve and cell markers have no ob-vious changes.Conclusions:The conditioned culture media supplemented with Y27632 have potential to effectively expand non-odonto-genic epithelial cells in vitro,and maintain the characteristics of stem cells,which is expected for tissue engineering tooth as seed cells.